To establish a highly efficient, sensitive, and accurate ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS/MS) method to determine the concentration of melatonin (MLT) in human plasma with heparin sodium.

The concentration of melatonin in plasma was measured using UHPLC-MS/MS. Plamsa sample preparation was done by protein precipitation using acetonitrile, its isotope melatonin-d4 (MLT-d4) was used as the internal standard (IS). Using UPLC BEH C₁₈ (2.1 mm×50.0 mm, 1.7 μm) reverse phase chromatography column, gradient elution was performed with a mobile phase consisting of acetonitrile-water (0.05 mM Ammonium fluoride) at a flow rate of 0.4 mL·min^-1 for 3 min. MLT was monitored using positive electrospray triple quadrupole mass spectrometer via multiple reaction monitoring (MRM) mode. The monitored transitions were set at m/z 233.0→174.0 for MLT. Moreover, the IS was set at m/z 237.2→178.2. This method was applied in a single-center, randomized, open-label, single oral dose pharmacokinetic trial, with 12 subjects in each group for fasting and after meals, administered a dose of 2 g (containing 4 mg of melatonin). Statistical analysis of pharmacokinetic parameters C_max (ΔC_max), AUC_0-t (ΔAUC_0-t), and AUC_0-∞ (ΔAUC_0-∞) using WinNonlin version 8.3 or above or SAS version 9.4 or above.

The achieved lower limit of quantification was 50.00 pg·mL^-1. The validated method had an excellent linearity in the range of 50.00-50 000.00 pg·mL^-1 for MLT y=2.00x+1.12×10^-4 (r²>0.999 4). The results of the intra-batch and inter-batch precision showed that the relative standard deviation (RSD) at different concentrations did not exceed 5.77%, and the accuracy was between 92.11% - 102.01% . The extraction recoveries of low, medium and high quality control samples of MLT were 103.35%, 113.75% and 131.21%, respectively. The coefficient of variation was 12.13%. No significant matrix effect was found. Through the pharmacokinetic study of 12 healthy humans, it was found for the first time that after taking the same dose of melatonin, the postprandial C_max was about 50% of that of fasting, but the AUC was very similar, indicating that food might affect the absorption and metabolism of melatonin in the body.

This method has strong specificity, good precision and high sensitivity, which can quickly and accurately determine the content of melatonin in human plasma with heparin sodium, and is used for the detection of human blood drug concentration and pharmacokinetics of melatonin.

To analyze the clinical effect of dapagliflozin tables combined with semaglutide injection in the treatment of patients with type 2 diabetes mellitus (T2DM) accompanied by abdominal obesity.

Patients with T2DM accompanied by abdominal obesity were divided into the control group and the treatment group by the random number table method. The control group took dapagliflozin tables orally once a day, 10 mg each time. The treatment group was hypodermic injection with semaglutide injection once a week on the basis of the control group, 0.5 mg each time. Both groups were treated for 3 months. The lipid metabolism, glucose metabolism, body mass index (BMI), waist circumference(WC), serological indicators, adipokines and adverse drug reactions were compared between the 2 groups.

In this study, 84 cases were enrolled in the treatment group and 83 cases in the control group, and 3 cases fell off in both groups. Finally, 81 cases were enrolled in the treatment group and 80 cases in the control group. After treatment, the low-density lipoprotein cholesterol (LDL-C) in the treatment group and the control group were (3.21±0.43) and (3.57±0.54) mmol·L^-1, respectively, and the glycated hemoglobin (HbAlc) were (5.42±0.67)% and (6.95±0.75)%, respectively; the homeostasis model assessment of insulin resistance (HOMA-IR) were (2.02±0.37) and (2.45±0.48) respectively, the BMI were (26.02±2.11) and (27.12±2.38) kg·m^-2, respectively, and the WC were (90.25±8.17) and (95.07±9.14) cm, respectively; insulin-like growth factor (IGF) were (154.31±15.68) and (165.23±17.08) mmol·L^-1, respectively, and asprosin (ASP) were (1.20±0.31) and (1.37±0.38) μg·L^-1, respectively; the spexin (SPX) were (0.78±0.19) and (0.65±0.14) ng·mL^-1, respectively, while the difference of above indexes in treatment and control groups were all statistically significant (all P<0.05). The adverse drug reaction incidence in treatment and control groups were 8.64% and 5.00%, respectively, showing no statistically significant difference (P>0.05).

Dapagliflozin tables combined with semaglutide injection can effectively regulate the glycolipid metabolism of patients with type 2 diabetes mellitus (T2DM) accompanied by abdominal obesity, reduce BMI and WC, regulate adipokines, inhibit the expression of serum IGF, and with reliable safety.

To investigate the off-label drug use of escitalopram in children and adolescents, and to provide a reference for rational clinical medication.

Collected prescription data of children and adolescents (<18 years) receiving antidepressant treatment, including gender, age, clinical diagnosis, drug name, dosage, frequency and route of administration. Based on domestic drug labeling, we assessed whether escitalopram was used off-label, and further conducted evidence-based searches regarding its off-label drug use. A literature search was conducted using the keywords escitalopram, off-label use, evidence-based medicine, children and adolescents across multiple electronic databases, including PubMed, Embase, The Cochrane Library, Web of Science, China National Knowledge Infrastructure (CNKI), Wanfang and VIP.

This study included a total of 156 504 outpatients, with escitalopram prescribed to 9 798, the off-label use rate was 6.26%. An analysis of the off-label use of escitalopram was conducted based on 22 articles, including the original drug instructions, evidence-based databases and related medical literature, the results demonstrated that escitalopram was supported by evidence for the use of both depression and generalized anxiety disorder in children and adolescents.

Escitalopram has preliminary evidence-based support for its use in children and adolescent patients, it is an off-label application. Therefore, clinical practice should strengthen the supervision and management of off-label use of escitalopram to ensure medication safety for patients.

To develop and validate a simple scoring system for predicting the risk of infection after acute myocardial infarction (AMI) in patients undergoing percutaneous coronary intervention (PCI).

Continuous patients with ST-segment elevation myocardial infarction (STEMI) who received PCI in our hospital were divided into training set and validation set using a cohort approach. All patients received oral rosuvastatin calcium tablets 10-20 mg per day combined with evolocumab injection therapy 140 mg. The primary endpoint was post-AMI infection during hospitalization, while all-cause mortality and major adverse cardiovascular events (MACE) were considered secondary endpoints. The area under the curve (AUC) and the calibration between predicted and observed infection risks were calculated. A backward stepwise logistic regression model was selected to develop the risk score.

A total of 359 patients were included, with 217 in the training set and 142 in the validation set, and no patient dropout occurred during the study. A 24-point risk scoring system was developed, with infection risks ranging from 0.70% to 99.60% in patients with the lowest and highest scores, respectively. Seven variables including age, Killip class, insulin use, white blood cell count, serum albumin, diuretics and transfemoral approach were incorporated into the model. The model achieved equally high discrimination in both the development and validation cohorts (C-statistic: 0.85) and demonstrated good calibration in both datasets. In both development and validation cohorts, the incidence of post-myocardial infarction infection increased steadily across risk score groups. Additionally, this risk score exhibited good predictive ability for infection, in-hospital all-cause mortality.

This study established a simple bedside tool, a risk scoring system for assessing the risk of infection and other in-hospital outcomes in STEMI patients undergoing PCI. Clinicians can utilize this risk scoring system to evaluate infection risk and make evidence-based decisions accordingly.

To compare the clinical efficacy and safety of labetalol injection and nifedipine controlled-release tablets combined with magnesium sulfate injection and calcium carbonate D3 tablets in the treatment of preeclampsia.

The patients with preeclampsia who were hospitalized in the obstetrics department of our hospital were included as the research objects, and the patients were divided into group A, group B and group C according to the medication plan. Group A was treated with labetalol hydrochloride injection 100-200 mg, intravenous drip, 1.5-3.5 mg per minute, once a day combined with 25% magnesium sulfate injection 60 mL, intravenous drip, 1.55-2.0 g per hour, 25 g per day and calcium carbonate D3 tablets, take orally, 300 mg each time, twice a day; Group B was treated with nifedipine controlled-release tablets orally, 30 mg each time, once a day, and the administration methods of magnesium sulfate injection, calcium carbonate D3 tablets were the same as those in group A; Group C was treated with labetalol hydrochloride injection, nifedipine controlled-release tablets, magnesium sulfate injection and calcium carbonate D3 tablets, and the administration methods of them were the same as those in group A and group B. The clinical efficacy, blood pressure condition, heart and kidney function, psychological status, fetal hemodynamics, maternal and pregnancy outcome of patients in 3 groups were compared, and the safety was evaluated.

A total of 170 patients were enrolled, 90 patients in group A, 39 patiets in group B and 41 patients in group C. After treatment, the total effective rates of group A, Group B and Group C were 58.89% (53 cases/90 cases), 53.85% (21 cases / 39 cases) and 85.37% (35 cases / 41 cases), respectively, and the blood pressure recovery time were 3.00 (3.00,5.00), 3.00 (3.00,5.00), and 7.00 (3.00,9.00) d, respectively. The total effective rate of group C was statistically significantly higher than that of group A and group B (all P<0.05), and the recovery time of blood pressure was statistically significantly shorter than that of group A and group B (all P<0.05). After treatment, the systolic blood pressures of group A, group B and group C were (145.60±5.15), (143.72±4.86) and (137.24±5.07) mmHg, respectively; the diastolic blood pressures were (97.83±6.56), (96.72±7.00) and (86.80±4.87) mmHg, respectively; Hamilton anxiety scale scores were 15.00 (13.00,17.00), 15.00 (13.50,17.50) and 12.00 (11.00,14.00) points, respectively; the Hamilton depression scale scores were 14.00 (12.00,15.00), 14.00 (12.00,15.00) and 10.00 (9.00,12.00) points, respectively; and the above indicators in group C were all lower than those of group A and group B (all P<0.05). After treatment, the B-type natriuretic peptide levels in group A, group B and group C were 35.00 (21.00,56.00), 42.00 (22.50,90.50) and 24.00 (13.00,60.00) pg·L^-1, respectively; the the urinary protein levels were 0.35 (0.14,0.57), 0.24 (0.15, 0.39) and 0.14 (0.06,0.34) g, respectively; and the B-type natriuretic peptide level of group C was statistically significantly lower than that of group B (P<0.05), and the urinary protein level was statistically significantly lower than that of group A (P<0.05). After treatment, the diastolic flow velocity ratio in group A, Group B and group C were 2.15 (1.92, 2.24), 2.04 (1.86, 2.23), 2.16 (2.02, 2.25), respectively; the flow resistance index were 0.53 (0.48, 0.55), 0.51 (0.46, 0.56), 0.54 (0.51, 0.58), respectively; the flow pulse index were 0.77 (0.66, 0.83), 0.69 (0.63, 0.80), 0.77 (0.67, 0.81), respectively; the incidence rates of adverse drug reactions were 13.33% (12 cases / 90 cases), 12.82% (5 cases / 39 cases) and 12.20% (5 cases / 41 cases), respectively; and there were no statistically significant differences in the above indicators and pregnancy outcome among the three groups (all P>0.05).

The therapy with labetalol, nifedipine, magnesium sulfate and calcium in PE is better, which can improve the blood pressure level, psychological status, heart and kidney function, maternal and infant outcomes, regulate fetal hemodynamics, and will not cause serious adverse reactions.

To analyze the clinical efficacy and safety of polyenophosphatidylcholine (PPC) in combination with tenofovir tablets in the treatment of patients with hepatitis B combined with nonalcoholic steatohepatitis (NASH).

The patients with hepatitis B complicated with NASH in our hospital were divided into treatment group and control group by random number table method. The control group was treated with tenofovir tablets, 25 mg qd, taken during meals. The treatment group was treated with PPC on the basis of the control group. The initial dose was 456 mg tid, taken with warm boiled water, and the daily dose could not exceed 1 368 mg. After 12 weeks of treatment, the dose was reduced to 228 mg tid, according to the immune response, and the dose was maintained. Both groups were treated for 24 weeks, and the treatment group was followed up to 48 weeks after stopping PPC. The negative conversion rates of hepatitis B virus surface antigen (HBsAg) and hepatitis B virus e antigen (HBeAg), hepatitis B virus DNA (HBV-DNA) load, the levels of costimulatory molecule B7 homolog 1 (B7-H1), transforming growth factor β1 (TGFβ1), Golgi transmembrane glycoprotein 73 (GP73), chitinase 3-like protein 1 (CHI3L1) and prothrombin activity (PTA) were compared between the two groups, and the safety was evaluated.

A total of 95 cases were enrolled and 92 cases were included in statistical analysis, including 46 cases in the treatment group and 46 cases in the control group. After treatment, the total effective rates of the treatment group and the control group were 97.83% (45 cases / 46 cases) and 84.78% (39 cases / 46 cases), respectively, and the difference was statistically significant (P<0.05). After 24 weeks of treatment, the HBV-DNA load of the treatment group and the control group were 80.43% and 60.87%, respectively; the HBsAg negative conversion rates were 28.26% and 10.87%, respectively; and the HBeAg negative conversion rates were 30.43% and 13.04%, respectively. The above indicators in the treatment group were statistically significantly different from those in the control group (all P<0.05). After 48 weeks of treatment, the HBV-DNA load of the treatment group and the control group were 89.13% and 69.57%, respectively; the HBsAg negative conversion rates were 50.00% and 28.26%, respectively; and the HBeAg negative conversion rates were 50.00% and 26.09%, respectively. The above indicators in the treatment group were statistically significantlly different from those in the control group (all P<0.05 ). After treatment, the levels of B7-H1 in the treatment group and the control group were 0.40 (0.30, 0.40) and 0.60 (0.50, 0.70) ng·mL^-1, the levels of TGFβ1 were (269.02±24.36) and (287.64±27.39) ng·mL^-1, respectively; and the levels of GP73 were (76.24±8.95) and (87.22±10.03) ng·mL^-1, respectively; PTA were (80.02 ± 12.55)% and (65.24±10.24)%, respectively. The treatment group compared with the control group, the differences were statistically significant (all P<0.05). The main adverse drug reactions during treatment in the treatment group and the control group were nausea and vomiting, headache, rash and abdominal distension. The total incidence of adverse drug reactions in the treatment group was 10.87% (5 cases / 46 cases), which was 13.04% (6 cases / 46 cases) in the control group, and the difference had no statistically significant (P>0.05).

The combined application of PPC and tenofovir tablets in the treatment of hepatitis B patients with NASH can reduce the levels of B7-H1 and TGFβ1, improve the liver function of the body, and has good safety.

To investigate the association between drug resistant epilepsy (DRE) and single nucleotide polymorphisms (SNPs) in whole-exome sequencing (WES).

Patients with epilepsy, who visited in our hospital, were enrolled in this study and divided into a drug-resistant group and a monotherapy-effective group based on treatment outcomes. Baseline demographic characteristics, medical history and laboratory test results were collected. Seizure outcomes were observed after more than 12 months under adequate treatment. WES was performed on 12 extreme cases (6 patients with resistant to three antiepileptic drugs vs. 6 patients effectively treated with monotherapy and undergoing dose reduction) to identify candidate SNPs. These candidate SNPs were then validated in the larger cohort to explore the association with DRE.

A tota of 560 patients were enrolled, 153 were DRE group and 407 were monotherapy-effective group. The median age in the monotherapy-effective group and the drug-resistant group was 19 years old and 25 years old, respectively; the median weight were 50 kg and 55 kg, respectively. All these differences between the two groups were statistically significant (all P<0.05); the proportion of structural etiology was 55.56% in the drug-resistant group and 45.46% in the monotherapy-effective group; the proportion of infectious etiology were 3.92% and 2.21%; the proportion of immune etiology were 3.27% and 1.23%; the proportion of metabolic and genetic etiology were 5.88% and 11.30%; the proportion of dual etiologies were 1.96% and 1.23%; the proportion of patients with impaired consciousness were 56.21% and 39.07%; the proportion of second-generation antiseizure medication as initial medication regimen were 50.98% and 65.46%; the proportion of malformation of cortical development were 10.46% and 2.95%; the proportion of hippocampal sclerosis were 15.03% and 9.58%; the proportion of hypoxic-ischemic encephalopathy were 3.92% and 1.47%; the proportion of vascular malformation were 4.58% and 3.93%; the proportion of traumatic brain injury were 1.31% and 3.44%; the proportion with brain tumor were 3.92% and 2.21%; the proportion with dual pathologies were 1.96% and 0.98%; the proportion of abnormal nonepileptogenic were 23.53% and 29.48%. Baseline information showed statistically significant differences in age, weight, etiology, presence of impaired awareness, initial medication regimen and neuroimaging results (P<0.05, P<0.001). 102 candidate SNPs were selected for genotyping and association analysis in larger cohort including 560 patients. 5 SNPs were identified to be significantly associated with DRE. Multivariate logistic regression incorporating both clinical and genetic factors revealed that hippocampal sclerosis, malformation of cortical development, hypoxic-ischemic, presence of impaired awareness, as well as the SCN2A rs17183814 GG and SPATA31 rs1919128 GA genotypes were risk factors for drug resistance (SCN2A rs17183814 GG: P=0.027; SPATA31 rs1919128 GA: P=0.014).

SCN2A rs17183814 and SPATA31 rs1919128 were found to be associated with drug resistance. Carriers of the GG genotypes of SCN2A rs17183814 and GA genotype of SPATA31 rs1919128 are more likely to develop DRE.

To establish and validate a high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for simultaneously determining the serum concentration of lamotrigine(LTG), levetiracetam(LEV), oxcarbazepine(OXC) and monohydroxycarbazepine(MHD), which is applied for clinical blood concentration detection.

A simple one-step precipitation method was used for the serum sample pretreatment, and stable isotopes of two antiepileptic drugs (OXC-D4, LEV-D6) as the internal standard. The mobile phases were aqueous solution of 0.1% formic acid and 5 mM ammonium acetate(A) and methyl alcohol(B), and the gradient elution was performed at the speed of 0.6 mL·min^-1 for 6 min on Phenomenex C₁₈(100.0 mm×4.6 mm, 3.0 μm). The column temperature was 40 ℃. Positive ion scan analysis was carried out in multi-reaction monitoring mode, using electrospray ion source. The specificity, standard curve and lower limit of quantification (LLOQ), accuracy and precision, extraction recovery, matrix effect and stability were evaluated. The patients treated with LRG, LEV or OXC in hospital were included as the research object. Blood samples were collected after the blood concentration reached a steady state and the serum concentration was detected by the method developed in this study.

The method demonstrated good specificity. For LTG, LEV, OXC and MHD, outstanding linearity was observed in the ranges of (0.20-25.00), (0.39-50.00), (0.04-5.00), (0.31-40.00) μg·mL^-1, respectively. The linear equation of LTG was y=0.99x+0.02 (r=0.999 2), LEV was y=0.55x+0.01 (r=0.999 4), OXC was y=4.35x+0.04 (r=0.998 9), and MHD was y=0.56x+0.01 (r=0.998 7). The LLOQs were 0.20, 0.39, 0.04, 0.31 μg·mL^-1, respectively. Acceptable accuracy as well as intra-batch and inter-batch precision were achieved; relative error (RE) and relative standard deviation (RSD) were all <15.00%. The extraction recoveries reached the standard (88.56%~112.18%, RSD<15.00%), without significant matrix effects and carryover effects. The serum samples of LTG, LEV, OXC, and MHD exhibited stability under the following conditions: storage at -20 ℃ for 14 days, three freeze-thaw cycles (-20 ℃ to room temperature), room temperature exposure for 3 hours, and post-treatment storage in the autosampler (4 ℃) for 24 hours, with all RSDs ≤10.72%. The analysis of 145 clinical samples showed that the linear range could meet the clinical needs.

This study successfully established a HPLC-MS/MS method to simultaneously quantify the serum concentration of LTG, LEV, OXC and MHD. The method is simple, economical accurate and efficient, which has promotional value.

To explore the potential role of platelet factor 4 (PF4) in Alzheimer’s disease (AD) by network pharmacology and bioinformatics analysis.

PubChem and SwissTargetPrediction databases were used to collect PF4 related targets, GeneCards and DisGeNET databases were used to collect AD targets; the protein-protein interaction (PPI) network was constructed by using STRING database to obtain the protein-protein interaction of common genes. Gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis of common targets were performed using DAVID database. The VascularSingleCells database was used to query the expression of PF4 in mouse cerebrovascular-related cells, and the AlzData database was used to analyze the verification of PF4 in the data set of known AD patients.

A total of 102 PF4-related targets, 2 403 AD targets and 51 common targets were obtained. Enrichment analysis showed that the common argets were mainly involved in cancer pathways, neutrophil extracellular trap formation, Interleukin-17 (IL-17) signaling pathway, platelet activation and relaxin signaling pathway. Analysis of cerebrovascular single cell data showed that PF4 was only expressed in microglia. The results of HPA database analysis showed that PF4 was mainly expressed in the cerebral cortex, and a small part was expressed in the amygdala.

PF4 may interfere with AD by improving neuronal viability, oxidative stress and reducing inflammatory response through neutrophil extracellular trapping network formation, IL-17 signaling and other pathways.